Gut microbiome community analysis is used to understand many diseases like inflammatory bowel disease, obesity, and diabetes. Sampling methods are an important consideration for human microbiome ...research, yet are not emphasized in many studies. In this study, we demonstrate that the preparation, handling, and storage of human faeces are critical processes that alter the outcomes of downstream DNA-based bacterial community analyses via qPCR. We found that stool subsampling resulted in large variability of gut microbiome data due to different microenvironments harbouring various taxa within an individual stool. However, we reduced intra-sample variability by homogenizing the entire stool sample in liquid nitrogen and subsampling from the resulting crushed powder prior to DNA extraction. We experimentally determined that the bacterial taxa varied with room temperature storage beyond 15 minutes and beyond three days storage in a domestic frost-free freezer. While freeze thawing only had an effect on bacterial taxa abundance beyond four cycles, the use of samples stored in RNAlater should be avoided as overall DNA yields were reduced as well as the detection of bacterial taxa. Overall we provide solutions for processing and storing human stool samples that reduce variability of microbiome data. We recommend that stool is frozen within 15 minutes of being defecated, stored in a domestic frost-free freezer for less than three days, and homogenized prior to DNA extraction. Adoption of these simple protocols will have a significant and positive impact on future human microbiome research.
•CMC coating extended cold storage life and maintained Kinnow mandarin fruit quality.•CMC retarded PME and cellulase enzyme activities up to 60 days cold stored fruit.•CMC coating-maintained SSC, TA, ...vitamin C, sensory traits up to 60-day stored fruit.
Edible coatings are safer for human health and environment compared to synthetic waxes. The aim of this study was to assess the influence of different edible coatings on extending cold storage life and maintaining fruit quality of ‘Kinnow’ mandarin (Citrus nobilis L. x Citrus deliciosa L.). Kinnow fruit coating treatments included 1.0, 1.5 and 2.0 g L−1 carboxy methylcellulose (CMC), 0.5, 1.0 and 1.5 g L−1 chitosan and 5, 10 and 15 g L−1 beeswax were contrasted to the uncoated (control) fruit. The coated and uncoated fruit were kept under cold storage conditions (5–7 °C and 90–95% RH) up to 75 days. The fruit were sampled following 30, 45, 60 and 75 days cold storage and evaluated for different quality parameters and scanning electron microscopy of the peel surface. The results indicate variability among the different polymer coatings for weight loss, spoilage loss, juice content, firmness, pectin content, the activity of pectin methylesterase and cellulase, soluble solids concentration (SSC), titratable acidity (TA), ascorbic acid content, total carotenoids content and organoleptic sensory attributes. Improved fruit gloss was observed in all coatings treatments, however, with the extension of the cold storage period, the fruit coated with chitosan and beeswax showed outward ruptures on the rind. Amongst all, the coatings tested, CMC (2.0 g L-1) coating was the most effective in maintaining various fruit quality parameters such as SSC, TA, vitamin C, sensory attributes and retarding the activities of cell wall degrading enzymes such as pectin methylesterase and cellulase up to 60 days in cold storage.
Abstract
The challenges facing domestic violence (DV) survivors has become the subject of a growing body of intersectional analysis; this paper presents a new intersectional framework for analysing ...DV policy and practice. Examining a partnership between women’s sector organizations and the criminal justice system in London, using interviews with professionals from the bodies, the paper offers an intersectional analysis of the implications, positive and negative, for both policy and practice. The complex interplay between women’s sector organizations and the criminal justice system demonstrate how the intersections of gender, class, ‘race’ and immigration status affect the way DV survivors are able to access and benefit from support, concluding that, to operate more effectively, DV support must consider all forms of oppression and violence affecting the lives of all their users.
The next galactic core-collapse supernova (CCSN) has already exploded, and its electromagnetic (EM) waves, neutrinos, and gravitational waves (GWs) may arrive at any moment. We present an extensive ...study on the potential sensitivity of prospective detection scenarios for GWs from CCSNe within 5 Mpc, using realistic noise at the predicted sensitivity of the Advanced LIGO and Advanced Virgo detectors for 2015, 2017, and 2019. We quantify the detectability of GWs from CCSNe within the Milky Way and Large Magellanic Cloud, for which there will be an observed neutrino burst. We also consider extreme GW emission scenarios for more distant CCSNe with an associated EM signature. We find that a three-detector network at design sensitivity will be able to detect neutrino-driven CCSN explosions out to ~5.5kpc, while rapidly rotating core collapse will be detectable out to the Large Magellanic Cloud at 50 kpc. Of the phenomenological models for extreme GW emission scenarios considered in this study, such as long-lived bar-mode instabilities and disk fragmentation instabilities, all models considered will be detectable out to M31 at 0.77 Mpc, while the most extreme models will be detectable out to M82 at 3.52 Mpc and beyond.
Abstract
British South Asian male child sexual abuse (CSA) survivors face a twofold problem: lack of research on this phenomenon and limited service provision. First, male CSA survivors have ...traditionally been marginalized in favour of female CSA survivors, a group that forms the basis of most academic research and to whom the majority of counselling and support services in the UK cater (Rapsey et al. 2020). The centrality of these female experiences, although crucial to understanding CSA survivor experiences in general, has eclipsed the specific narratives of male survivors. Second, despite the UK’s incredibly multicultural population, people from minoritized backgrounds still experience institutional forms of racism that are entrenched in service provision and research and that lead to racialized and Eurocentric perspectives and practices (Singh 2019). This article thus seeks to expand on the existing body of literature on male CSA in minoritized groups. It illuminates the experiences of male survivors from British South Asian communities, creating space for the much-needed voices of a critically under-researched group.
Cystic pancreatic lesions (CPLs) are increasingly detected by various imaging studies. Mucinous CPLs carry a risk of malignant transformation but this is often difficult to diagnose preoperatively. ...In a previous report of 10 suspected mucinous CPLs, the cellular yield of endoscopic ultrasonography (EUS)-guided cytology brushings was found to be superior to the yield from standard fine-needle aspiration (FNA). The aim of this prospective and blinded study was to compare the cytology yield of mucinous epithelium from brushing with FNA in suspected mucinous CPLs.
In total, 37 patients with 39 CPLs measuring at least 20 mm were enrolled between June 2006 and July 2008 for EUS-cytobrushing and EUS-FNA of CPLs. Demographic, clinical, EUS, cytopathologic, and surgical data were recorded whenever available. Yield of cytology brushings was compared with that of FNA. Procedure morbidity was evaluated after 30 days. The main outcome assessed was yield of intracellular mucin (ICM) on cytobrushing specimens compared with EUS-FNA for the diagnosis of suspected mucinous CPL.
Cytobrushings were more likely to detect ICM than the EUS-FNA method ( P = 0.001). In three patients with hypocellular FNA, dysplasia was found on cytology brushing and later confirmed by surgical pathology. Significant complications occurred in three patients (8 %): one postbrushing bleeding and two acute pancreatitis.
Cytology brushings are more likely to provide an adequate mucinous epithelium specimen than standard FNA and could aid the diagnosis of CPLs in a selective group of patients.
The gastrointestinal tract is enveloped by concentric and orthogonally aligned layers of smooth muscle; however, an understanding of the mechanisms by which these muscles become patterned and aligned ...in the embryo has been lacking. We find that Hedgehog acts through Bmp to delineate the position of the circumferentially oriented inner muscle layer, whereas localized Bmp inhibition is critical for allowing formation of the later-forming, longitudinally oriented outer layer. Because the layers form at different developmental stages, the muscle cells are exposed to unique mechanical stimuli that direct their alignments. Differential growth within the early gut tube generates residual strains that orient the first layer circumferentially, and when formed, the spontaneous contractions of this layer align the second layer longitudinally. Our data link morphogen-based patterning to mechanically controlled smooth muscle cell alignment and provide a mechanistic context for potentially understanding smooth muscle organization in a wide variety of tubular organs.
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•Gut smooth muscle is patterned by molecular signals and aligned by mechanical forces•Hedgehog and Bmp signaling control spatiotemporal differentiation of muscle layers•Differential growth generates static strain that aligns the inner circumferential layer•Cyclic contractions of the circumferential layer orient the longitudinal layer
Gut smooth muscle patterning entails mechanical strain that orients the first layer circumferentially, and when formed, it spontaneously contracts to align the second layer longitudinally.
Parvalbumin inhibitory interneurons (PVIs) are crucial for maintaining proper excitatory/inhibitory balance and high-frequency neuronal synchronization. Their activity supports critical developmental ...trajectories, sensory and cognitive processing, and social behavior. Despite heterogeneity in the etiology across schizophrenia and autism spectrum disorder, PVI circuits are altered in these psychiatric disorders. Identifying mechanism(s) underlying PVI deficits is essential to establish treatments targeting in particular cognition. On the basis of published and new data, we propose oxidative stress as a common pathological mechanism leading to PVI impairment in schizophrenia and some forms of autism. A series of animal models carrying genetic and/or environmental risks relevant to diverse etiological aspects of these disorders show PVI deficits to be all accompanied by oxidative stress in the anterior cingulate cortex. Specifically, oxidative stress is negatively correlated with the integrity of PVIs and the extracellular perineuronal net enwrapping these interneurons. Oxidative stress may result from dysregulation of systems typically affected in schizophrenia, including glutamatergic, dopaminergic, immune and antioxidant signaling. As convergent end point, redox dysregulation has successfully been targeted to protect PVIs with antioxidants/redox regulators across several animal models. This opens up new perspectives for the use of antioxidant treatments to be applied to at-risk individuals, in close temporal proximity to environmental impacts known to induce oxidative stress.
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